摘要
目的观察乌司他丁（ulinastatin，UTI）对急性硫化氢（H2S）中毒大鼠脑损伤后水通道蛋白4（aquaporin4，AQP4）及环氧合酶-2（cyclooxygenase-2，COX-2）表达改变的影响，探讨H2S中毒脑损伤的机制和UTI的保护效应。方法将清洁级SD大鼠96只随机分为正常对照组（8只）、UTI对照组（8只）、H2S染毒组（40只）和UTI干预组（40只）。正常对照组和UTI对照组大鼠吸入与H2S同流量的空气1h，UTI对照组大鼠立即腹腔注射UTI（10万单位／kg），2h后正常对照组和UTI对照组大鼠均处死。H2S染毒组和UTI干预组大鼠在染毒柜内吸入浓度为284mg／m，的H2S 1h，UTI干预组立即腹腔注射UTI（10万单位／妇），两组大鼠分别于2、6、12、24、48h观察大鼠行为改变，并分别在各时间点处死，留取脑组织。RT-PCR法检测脑组织中AQP4、COX-2、NSE mRNA的表达，免疫组化Streptavidin-perosidase法测定AQP4、COX-2、神经元特异性稀醇化酶（NSE）蛋白表达。光学显微镜下观察各组脑组织病理学改变。结果H2S染毒组大鼠脑组织神经细胞水肿、变性，炎症细胞灶性浸润，毛细血管增生、扩张。与正常对照组比较，H2S染毒组各时间点NSE mRNA和蛋白表达明显增加，差异有统计学意义（P〈0.01）。与正常对照组比较，H2S染毒组各时间点AQP4及COX-2 mRNA和蛋白表达明显增加，差异有统计学意义（P〈0．01）。与H2S染毒组比较，6、12、24、48h UTI干预组大鼠脑组织中NSE mRNA和蛋白表达明显降低，差异有统计学意义（P〈0.01）。与H2S染毒组比较，UTI干预组6、12、24、48h时AQP4及COX-2 mRNA和蛋白表达明显降低，差异有统计学意义（P〈0．01）。结论急性H2S中毒脑损伤的发生与AQP4、COX-2异常表达有关，UTI干预能降低H2S中毒后脑组织中AQP4、COX-2水平，减轻脑损伤的程度。
Objective To observe the effect of Ulinastatin （UTI） in the dynamic changes of aquaporin 4 （AQP4） and cyclooxygenase-2 （COX-2） in the brain tissue injury of acute hydrogen sulfide （H2S） intoxicated, to explore the Mechanism of brain tissue injury of acute H2S-intoxicated and the protection effect of UTI. Methods A total of 96 SD rats of clean grade were divided randomly into four groups ： normal control group （NS group, n= 8）, UTI control group （UTI group, n=8 ）, H2S-intoxicated model group （H2S group, n=40）, UTI treatment group （H2S＋UTI group, n=40）. The H2S group and H2S＋UTI group were exposed to H2S（284 mg/m^3） by inhalation for l h, then H2S＋UTI group was intraperitoneally exposed to UTI at the close of 105 U/kg for 2 h, H2S group and HzS＋UTI group were sacrificed at 2,6,12,24 and 48 h after exposure, respectively. Remove the brain tissue, observe the rats behavioral changes at each time points. The mRNA expression of AQP4 ,COX-2 and NSE in the brain tissue were measured by RT-PCR method, and the protein expression of AQP4,COX-2 and NSE in the brain tissue were detected by immunohistoehemieal Streptavidin-perosidase method. Pathological changes of brain tissue were observed by lightmieroscope. Results 1 ,Nerve cells in the H2S group rats had edema, degeneration, focal inflammatory cell infiltrate, capillary hyperplasia, expansion. Compared with NS group, the cerebral NSE mRNA and protein expression at each time point in H2S group after exposure were significantly increased （P〈0.01）. 2,Compared with NS group, the cerebral AQP4 and COX-2 mRNA and protein expression at each time point in H2S group after exposure were significantly increased （P〈0.01）. 3 ,The degree of brain damage was significantly decreased in H2S＋UTI group than that in H2S group. Compared with H2S group, the cerebral NSE mRNA and protein expression at 6, 12, 24 and 48 h in HzS＋UTI group after exposure were significantly decreased （P〈0.01）, no significantly difference at 2h （P〉0.05）. 4 ,Compared with the H2S group, the cerebral AQP4 and COX-2 mRNA and protein expression at 6, 12, 24 and 48 h in H2S＋UT! group after exposure were significantly decreased（P〈0.01 ）, slightly decreased at 2 h. Conclusion The mechanism of brain injury of acute hydrogen sulfide intoxicated associated with abnormal expression of the cerebral AQPg, COX-2 levels. Intervention of UTI can reduce the cerebral AQP4 and COX-2 levels after hydrogen sulfide intoxicated, reduce the degree of brain injury.
出处
《中华劳动卫生职业病杂志》
CAS
CSCD
2016年第3期166-172,共7页
Chinese Journal of Industrial Hygiene and Occupational Diseases
基金
浙江省中医药科技计划项目（2011ZA074）
浙江省中医药重点学科计划（2012-XK-A28）
浙江省“十二五”重点学科建设项目（2012-207）
浙江省医学创新学科建设项目（11-CX26）
